Lee Doh Young, Hah J Hun, Jeong Woo-Jin, Chung Eun-Jae, Kwon Tack-Kyun, Ahn Soon-Hyun, Sung Myung-Whun, Kwon Seong Keun
Department of Otorhinolaryngology-Head and Neck Surgery, Seoul National University Boramae Medical Center, Seoul, Korea.
Department of Otorhinolaryngology-Head and Neck Surgery, THANQ Seoul Thyroid-Head and Neck Surgery Center, Seoul, Korea.
Clin Exp Otorhinolaryngol. 2022 Nov;15(4):372-379. doi: 10.21053/ceo.2022.00150. Epub 2022 Nov 16.
We aimed to assess the genetic differences between cases of early-stage tongue cancer that were positive or negative for lymph node metastasis.
In total, 35 cases of tongue cancer with RNA sequencing data were enrolled in this study. The gene expression profile of the following two groups was compared: N0 group (T stage 1 or 2 with N0 stage) and N+ group (T stage 1 or 2 with N+ stage). Using the R and limma packages in the Bioconductor program, we extracted the differentially expressed genes (DEGs). Gene ontology and pathway enrichment analysis were performed using the Database for Annotation, Visualization and Integration Discovery (DAVID) online tool. Immune cell infiltration was analyzed using the CIBERSORT online program. Immunochemical staining of the cancer tissue was evaluated and The Cancer Genome Atlas (TCGA) data were analyzed to validate the identified DEGs.
No significant differences were found in the infiltration of 22 types of immune cells. Among a total of 51 identified DEGs, 14 genes were significantly upregulated, while 37 genes were significantly downregulated (P<0.01; fold change >2). Pathway analysis revealed significant associations with the arachidonic acid metabolism-related pathway, calcium signaling, and the muscle contraction pathway. The following DEGs were the most significantly different between the two groups: DEFB4A, SPRR2B, DEFB103B, SPRR2G, DEFB4B, and FAM25A. TCGA data showed that DEFB4A and DEFB103B were more highly expressed in the N0 group than in the N+ group, although the difference did not achieve statistical significance. Immunochemical staining of cancer tissue revealed significantly higher expression of defensin in the N0 group.
. Defensin (DEFB4A, DEFB103B, DEFB4B) may be a novel biomarker for early regional metastasis in T1/2 tongue cancer.
我们旨在评估早期舌癌病例中淋巴结转移呈阳性或阴性的基因差异。
本研究共纳入35例有RNA测序数据的舌癌病例。比较了以下两组的基因表达谱：N0组（T1或T2期且N0期）和N+组（T1或T2期且N+期）。使用生物导体程序中的R和limma软件包，我们提取了差异表达基因（DEG）。使用在线工具注释、可视化和综合发现数据库（DAVID）进行基因本体和通路富集分析。使用CIBERSORT在线程序分析免疫细胞浸润情况。对癌组织进行免疫化学染色评估，并分析癌症基因组图谱（TCGA）数据以验证鉴定出的DEG。
在22种免疫细胞的浸润方面未发现显著差异。在总共鉴定出的51个DEG中，14个基因显著上调，37个基因显著下调（P<0.01；变化倍数>2）。通路分析显示与花生四烯酸代谢相关通路、钙信号传导和肌肉收缩通路有显著关联。以下DEG在两组之间差异最为显著：DEFB4A、SPRR2B、DEFB103B、SPRR2G、DEFB4B和FAM25A。TCGA数据显示，DEFB4A和DEFB103B在N0组中的表达高于N+组，尽管差异未达到统计学显著性。癌组织的免疫化学染色显示N0组中防御素的表达显著更高。
防御素（DEFB4A、DEFB103B、DEFB4B）可能是T1/2期舌癌早期区域转移的一种新型生物标志物。
